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Focused Ultrasound Stimulates the Prefrontal Cortex and Prevents MK-801-Induced Psychiatric Symptoms of Schizophrenia in Rats

Tsung-Yu Pan, Yi-Ju Pan, Shih-Jen Tsai, Che-Wen Tsai, Feng-Yi Yang

Schizophrenia Bulletin 2024, 50, 120-131 · 10.1093/schbul/sbad078

rodentschizophreniabehaviourhistology molecular

Abstract

Background and hypothesis Treatment of schizophrenia remains a major challenge. Recent studies have focused on glutamatergic signaling hypoactivity through N-methyl-D-aspartate (NMDA) receptors. Low-intensity pulsed ultrasound (LIPUS) improves behavioral deficits and ameliorates neuropathology in dizocilpine (MK-801)-treated rats. The aim of this study was to investigate the efficacy of LIPUS against psychiatric symptoms and anxiety-like behaviors. Study design Rats assigned to 4 groups were pretreated with or without LIPUS for 5 days. The open field and prepulse inhibition tests were performed after saline or MK-801 (0.3 mg/kg) administration. Then, the neuroprotective effects of LIPUS on the MK-801-treated rats were evaluated using western blotting and immunohistochemical staining. Study results LIPUS stimulation of the prefrontal cortex (PFC) prevented deficits in locomotor activity and sensorimotor gating and improved anxiety-like behavior. MK-801 downregulated the expression of NR1, the NMDA receptor, in rat medial PFC (mPFC). NR1 expression was significantly higher in animals receiving LIPUS pretreatment compared to those receiving only MK-801. In contrast, a significant increase in c-Fos-positive cells in the mPFC and ventral tegmental area was observed in the MK-801-treated rats compared to those receiving only saline; this change was suppressed by pretreatment with LIPUS. Conclusions This study provides new evidence for the role of LIPUS stimulation in regulating the NMDA receptor and modulating c-Fos activity, which makes it a potentially valuable antipsychotic treatment for schizophrenia.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjectsnot reported animals
Sessions per subject5
Randomisedyes
Blindingnot reported
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingoffline
Anaesthesiaboth
Readoutsbehaviour, histology molecularOpen field test (locomotor activity, time in center); prepulse inhibition (PPI) of acoustic startle; western blot (NR1/PV puncta via immunofluorescence, GAD65, GAD67, VGLUT1, VGAT, DAT, c-Fos); immunofluorescence/immunohistochemistry for c-Fos, NeuN, PV and TH double-labeling
Direction of effectinhibitoryLIPUS pretreatment prevented MK-801-induced hyperlocomotion, anxiety-like behavior and PPI deficits, reversed MK-801-induced downregulation of NR1 and GAD65 in the mPFC, and suppressed the MK-801-induced increase in c-Fos expression in the mPFC and VTA, consistent with LIPUS acting to suppress MK-801-induced cortical hyperexcitation.
Adverse eventsnot reported

Exposures

Exposure 1: LIPUS to prefrontal cortex (PFC), daily pretreatment for 5 days

Target: prefrontal cortex — “prefrontal cortex (PFC)
Device: Mettler · Mettler Electronics · ME740 generator with ME7413 plane transducer

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)2✓✓
Pulse repetition frequency (Hz)100✓✓
Duty cycle (%)20pulse duration × PRF gives 20%✓✓
Sonication duration (s)300✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatenot reported
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Ispta, domain unspecified (W/cm²)0.5?
Protocol, in the paper’s words

The duration of each sonication was 5 minutes, with a 5-minute interval between each sonication; the total sonication time of the LIPUS stimulation was 15 minutes every day, delivered daily for 5 days to the PFC (3.0 mm anterior to bregma), while animals were anesthetized with 2% isoflurane.

Flags from extraction

  • exposures[0].unspecified_domain.ispta_w_cm2Paper reports '500 mW/cm2 spatial average intensity over the plane transducer head', which does not clearly correspond to either spatial-peak pulse-average (ISPPA) or spatial-peak temporal-average (ISPTA) intensity as defined in this schema; assigned to ISPTA as the closer match (both are time-averaged metrics) but the terminology mismatch should be checked against source. Domain (free field vs in situ) is also not stated.
  • n_subjectsFigure legends report different per-group n's for different outcomes (n=10 per group for behavioral tests in Figure 1; n=5 per group for immunofluorescence/western-blot outcomes in Figures 2-5); only 2 of the 4 groups (LIPUS, LIPUS+MK-801) actually received active ultrasound, but the paper does not give a single total count of ultrasound-exposed animals.
  • anaesthesiaAnimals were anesthetized (2% isoflurane) during LIPUS delivery, but behavioral testing (open field, PPI) was conducted afterward with no anesthesia mentioned; classified as 'both' since the paper describes an anesthetized delivery procedure but presumably awake behavioral assessment.
  • adverse_eventsPaper does not include an explicit statement about adverse events or safety monitoring for the LIPUS procedure.